"Chimeric enzymes enhance treatment potential for globoid cell leukodystrophy through hematopoietic stem cell gene therapy" and GR 2019-12369357 (final report)
Description
"Chimeric enzymes enhance treatment potential for globoid cell leukodystrophy through hematopoietic stem cell gene therapy" - PMID: 40988335 - DOI: 10.1016/j.ymthe.2025.09.030 Globoid cell leukodystrophy (GLD) is a fatal lysosomal storage disorder caused by a deficiency in the β-galactosylceramidase (GALC) enzyme, leading to severe demyelination and neurodegeneration, and often death before the age of 2 years. Hematopoietic stem/progenitor cell transplantation (HSPC-T) has limited efficacy due to inadequate GALC delivery to the central (CNS) and peripheral nervous systems (PNS) and associated risks. In vivo gene therapy (GT) using adeno-associated viral vectors shows promise, but safety concerns persist. This research presents a strategy using lentiviral (LV) vector-mediated ex vivo HSPC-GT with a chimeric GALC enzyme that incorporates peptides from α-L-iduronidase (IDUA) and apolipoprotein E II (APO) to enhance expression and blood-brain barrier penetration. The chimeric IDUAsp.GALC.APO enzyme exhibited superior production and secretion compared to native GALC and previous chimeric variants in LV-transduced HSPCs, resulting in improved cross-correction and normalization of GALC activity in GLD neural cells. Proof-of-concept studies demonstrated effective enzyme production, secretion, and cross-correction capability of macrophages from GLD patients. In vivo results showed stable gene marking, sustained enzyme production, and efficient delivery of the chimeric GALC in affected organs, including the CNS and PNS. These findings highlight the potential of HSPC-GT using chimeric GALC enzymes as an innovative therapeutic approach for treating GLD. Data related to the paper are available in the folder with the same name. "Enhancement of expression, bioavailability and cross-correction of chimeric GALC enzyme to refine gene therapy approaches for globoid cell leukodystrophy" - GR 2019-12369357 - Final Report In this project, we propose a lentiviral vector-mediated gene therapy with a chimeric GALC enzyme engineered for improved expression and blood-brain barrier penetration as therapeutic approach for GLD. Our findings demonstrate that the chimeric enzyme, when expressed in LV-transduced human HSPCs and GLD iPSC derived NPC progeny, exhibits enhanced production and secretion compared to native GALC and prior chimeric variants. This leads to significant cross-correction of GALC activity in GLD neural cells. In vivo studies in immunodeficient mice and a severe GLD mouse model confirmed stable gene marking and sustained enzyme production, emphasizing our strategy's potential for effective therapeutic delivery and cross-correction of affected tissues. The data related to the final report of GR 2019-12369357 are available in the folder with the same name.
Files
Institutions
Categories
Funding
Ministero della Salute
GR-2019-12369257
Telethon Foundation
TTAGD0222TT
European Leukodystrophy Association
ELA 2019-015I2
Fondazione Centro San Raffaele
FCSR-2019